H1R Signaling and Immune Deviation in EAE

EAE 中的 H1R 信号传导和免疫偏差

基本信息

批准号：
8602861
负责人：
CORY TEUSCHER
金额：
$ 32.49万
依托单位：
UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-01-19 至 2015-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8602861
关键词：
Adjuvant Affect Alleles Amines Amino Acids Arrestins Autoimmune Diseases Autoimmune Process Bordetella pertussis CD4 Positive T Lymphocytes Candidate Disease Gene Cell surface Cells Central Nervous System Diseases Chimeric Proteins Complement Data Development Disease Disease Resistance Disease susceptibility Endoplasmic Reticulum Enhancers Exhibits Experimental Autoimmune Encephalomyelitis Exposure to G Protein-Coupled Receptor Signaling G-Protein-Coupled Receptors GTP-Binding Proteins Gene Cluster Gene Transfer Techniques Genes Genetic Genetic Polymorphism Histamine Histamine H1 Receptors Hypersensitivity Immune Immune response In Vitro Inbred Strains Mice Inflammation Mediators Inflammatory Interferons Interleukin-17 Knowledge Lead Ligand Binding Link Linkage Disequilibrium Maps Mediating Mitogen-Activated Protein Kinases Modeling Molecular Chaperones Multiple Sclerosis Mus Mutation Pathway interactions Peptides Pertussis Toxin Phenotype Play Predisposition Process Production Proteins Receptor Activation Receptor Signaling Regulation Resistance Role SJL/J Mouse Signal Pathway Signal Transduction Signaling Protein Surface T-Lymphocyte Transfection Ursidae Family central nervous system demyelinating disorder cytokine forward genetics gene cloning human MAPK14 protein immune function in vivo loss of function mouse model mutant novel novel therapeutics oligodendrocyte-myelin glycoprotein overexpression protein activation protein folding protein transport public health relevance receptor expression research study restoration trafficking

项目摘要

DESCRIPTION (provided by applicant): Histamine is a potent mediator of inflammation and regulator of innate and adaptive immune responses, and autoimmune diseases such as experimental allergic encephalomyelitis (EAE), the principal autoimmune model of multiple sclerosis (MS). EAE is an inflammatory demyelinating disease of the central nervous system with a critical genetic component. We identified Bphs as a susceptibility locus for EAE and positional candidate gene cloning demonstrated Bphs to be the histamine H1 receptor (Hrh1/H1R). As in many autoimmune diseases, CD4+ T cells play a key role in MS and EAE through production of cytokines such as interferon-g and interleukin-17. T cell-specific expression of the susceptible (BphsS/H1RS) allele allowed for EAE development on a H1RKO background, while expression of the resistant (BphsR/H1RR) allele in the same fashion did not. H1R alleles differ by three amino acids in the third intracellular loop (H1RS: Pro263Val312Pro330; H1RR: Leu263Met312Ser330). This domain is associated with signal transduction, but may also be important in protein folding and trafficking, like the intracellular domains of other G-protein coupled receptors (GPCRs). In transfection studies, H1RR surface expression was substantially lower than H1RS, with H1RR exhibiting intracellular retention in the endoplasmic reticulum (ER). In Aim 1 we will dissect the mechanism(s) responsible for the ER-retention of the H1RR allele and whether pharmacological chaperones can restore H1RR surface expression and affect disease in vivo. Pharmacological chaperones restore surface expression and function of polymorphic GPCRs in vitro, and knowledge gained from the proposed in vivo experiments may have broad implications for GPCR-diseases. We also have evidence of a functional, complementary mechanism that can overcome the ER retention process in BphsR mice. Compilation of Hrh1 sequence data and Bphs data from ~100 inbred mouse strains, revealed strains that were phenotypically BphsS, but bear the H1RR allele. Preliminary mapping studies linked the locus to Hrh1 and because it corrects BphsR, we refer to this gene as Bphs-enhancer (Bphse). SJL/J mice exhibit another Hrh1-linked phenotype, spontaneous histamine sensitivity (Shs). We hypothesize that Bphse and Shs reside within a functional linkage disequilibrium (LD)-domain linked to Hrh1 and may contain genes related to H1R folding, trafficking, or signaling. In Aim 2 we will use a forward genetics approach to positionally clone and identify Bphse and Shs. H1R signaling through p38 mitogen activated protein kinase (MAPK) is important in EAE susceptibility. For many GPCRs sustained MAPK activation involves non-G-protein signaling through b-arrestins. Since we found no difference in G-protein activation of allelic H1R-Ga fusion proteins, this suggested that H1R signaling may also involve a novel non-G-protein b-arrestin pathway. Thus in Aim 3 we will examine whether non-G-protein signaling downstream of the H1R is required for EAE development. Non-G-protein signaling by GPCRs is an emerging paradigm and new pharmacologics are being developed to selectively target these pathways.

描述（由申请人提供）：组胺是炎症和适应性免疫反应调节剂的有效介质，以及自身免疫性疾病，例如实验性过敏性脑脊髓炎（EAE），这是多发性硬化症（MS）的主要自身免疫模型（MS）。 EAE是具有关键遗传成分的中枢神经系统的炎症性脱髓鞘疾病。我们将BPH识别为EAE和位置候选基因克隆的易感基因座，证明BPH是组胺H1受体（HRH1/H1R）。与许多自身免疫性疾病一样，CD4+ T细胞通过产生细胞因子（例如Interferon-g和interleukin-17）在MS和EAE中起关键作用。 T细胞特异性的表达（BPHSS/H1RS）等位基因允许在H1RKO背景下发育，而耐药性（BPHSR/H1RR）等位基因的表达方式却没有。 H1R等位基因在第三个细胞内环中的三个氨基酸（H1RS：Pro263VAL312PRO330; H1RR：LEU263MET312SER330）中的三个氨基酸不同。该结构域与信号转导有关，但在蛋白质折叠和运输中也可能很重要，例如其他G蛋白偶联受体（GPCR）的细胞内结构域。在转染研究中，H1RR表面表达大大低于H1R，H1RR在内质网（ER）中表现出细胞内保留。在AIM 1中，我们将剖析负责H1RR等位基因率的机制，以及药理学伴侣是否可以恢复H1RR表面表达并影响体内疾病。药理学伴侣在体外恢复了多态性GPCR的表面表达和功能，并且从所提出的体内实验中获得的知识可能对GPCR-脉络酶有广泛的影响。我们还有证据表明可以克服BPHSR小鼠的ER保留过程的功能互补机制。来自〜100个近交小鼠菌株的HRH1序列数据和BPHS数据的汇编显示，菌株在表型上是BPHS，但具有H1RR等位基因。初步的映射研究将轨迹与HRH1联系起来，并且由于它纠正了BPHSR，因此我们将此基因称为BPHS-ENHANCER（BPHSE）。 SJL/J小鼠表现出另一种HRH1连接的表型，自发组胺灵敏度（SHS）。我们假设BPHSE和SHS位于与HRH1相关的功能连锁不平衡（LD）域中，并且可能包含与H1R折叠，运输或信号传导有关的基因。在AIM 2中，我们将使用远期遗传学方法来定位克隆并识别BPHSE和SHS。通过p38有丝分裂原活化蛋白激酶（MAPK）的H1R信号在EAE易感性中很重要。对于许多GPCR，持续的MAPK激活涉及通过B- arrestin的非G蛋白信号传导。由于我们发现等位基因H1R-GA融合蛋白的G蛋白激活没有差异，因此这表明H1R信号传导也可能涉及一种新型的非G蛋白B- arrestin途径。因此，在AIM 3中，我们将检查H1R下游的非G蛋白信号传导是否需要EAE开发。 GPCRS的非G蛋白信号传导是一种新兴范式，正在开发新的药理学以选择性地靶向这些途径。